Pub Date : 2024-11-24DOI: 10.1016/j.ygcen.2024.114645
Yuta Tanizaki, Hongen Zhang, Yun-Bo Shi
Intestinal structure is drastically changed from fetal to adult form during postembryonic development, a period around birth in mammals. This process is regulated by thyroid hormone (T3) via its receptors, T3 receptor (TR) α and TRβ during anuran metamorphosis. Here, we used intestinal remodeling during Xenopus tropicalis metamorphosis, which serves as a model for human postembryonic development, to identify TR-bound genes and determine the relative contribution to target gene binding by TRα and TRβ. We first examined the localization of TRα and TRβ mRNA during metamorphosis in Xenopus tropicalis and found that TRα was broadly expressed in the intestinal tissues from premetamorphosis to the end of metamorphosis, while TRβ was expressed at low levels during premetamorphosis but was upregulated at the climax of metamorphosis when intestinal stem cells are formed and proliferate. Interestingly, both TR genes were co-expressed in different cell types, including stem cells. Chromatin immunoprecipitation (ChIP)-seq analyses of the intestine from wild type, TRα- or TRβ-knockout premetamorphic tadpoles treated with or without T3 for 18 h identified many TR-bound genes and revealed the effects of individual TR knockout on the binding of target genes by TR. We found that individual TR knockout reduced both the number of TR-bound genes and the extent of TR binding to target genes with TRα knockout had a much more dramatic effect than TRβ knockout. On the other hand, the TR-bound genes were largely common among the three genotypes. These findings suggest that both TRα and TRβ contribute to target binding with TRα having a bigger contribution in premetamorphic intestine.
{"title":"Complementary and additive functions of TRα and TRβ during intestinal remodeling as revealed by ChIP-Seq analysis on wild type and TR knockout animals.","authors":"Yuta Tanizaki, Hongen Zhang, Yun-Bo Shi","doi":"10.1016/j.ygcen.2024.114645","DOIUrl":"https://doi.org/10.1016/j.ygcen.2024.114645","url":null,"abstract":"<p><p>Intestinal structure is drastically changed from fetal to adult form during postembryonic development, a period around birth in mammals. This process is regulated by thyroid hormone (T3) via its receptors, T3 receptor (TR) α and TRβ during anuran metamorphosis. Here, we used intestinal remodeling during Xenopus tropicalis metamorphosis, which serves as a model for human postembryonic development, to identify TR-bound genes and determine the relative contribution to target gene binding by TRα and TRβ. We first examined the localization of TRα and TRβ mRNA during metamorphosis in Xenopus tropicalis and found that TRα was broadly expressed in the intestinal tissues from premetamorphosis to the end of metamorphosis, while TRβ was expressed at low levels during premetamorphosis but was upregulated at the climax of metamorphosis when intestinal stem cells are formed and proliferate. Interestingly, both TR genes were co-expressed in different cell types, including stem cells. Chromatin immunoprecipitation (ChIP)-seq analyses of the intestine from wild type, TRα- or TRβ-knockout premetamorphic tadpoles treated with or without T3 for 18 h identified many TR-bound genes and revealed the effects of individual TR knockout on the binding of target genes by TR. We found that individual TR knockout reduced both the number of TR-bound genes and the extent of TR binding to target genes with TRα knockout had a much more dramatic effect than TRβ knockout. On the other hand, the TR-bound genes were largely common among the three genotypes. These findings suggest that both TRα and TRβ contribute to target binding with TRα having a bigger contribution in premetamorphic intestine.</p>","PeriodicalId":12582,"journal":{"name":"General and comparative endocrinology","volume":" ","pages":"114645"},"PeriodicalIF":2.1,"publicationDate":"2024-11-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142727500","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-24DOI: 10.1016/j.ygcen.2024.114644
Kevin Pham, Madeline Choi, KayLene Yamada, Haruka Wada
Traits vary in their ability to recover from stressors and is dependent on the magnitude, duration, or type of stressor. One such stressor is circadian rhythm disruption stemming from artificial light at night (ALAN). Circadian rhythm disruption may lead to long-term physiological consequences; however, the capacity in which individuals respond to recovery and display stress resilience is not known. Here, we exposed zebra finches (Taeniopygia castanotis) to constant light (24L:0D) or a regular light/dark cycle (14L:10D) for 23 days and a recovery period for 12 days. We measured body mass, corticosterone, and glucose levels before, during, and after constant light exposure and relative protein abundance of glucocorticoid receptors at death. In light-exposed birds, body mass significantly increased over time compared to birds under a regular light/dark cycle, but a 12-day recovery period reversed this increase. Baseline levels of circulating glucose decreased in light-exposed birds compared to controls, but significantly increased after the 12-day recovery period. In contrast, the glucose stress response did not show a similar recovery trend as body mass or baseline glucose in light-exposed birds. We did not detect any changes in baseline corticosterone or stress reactivity in both groups throughout the experiment. Lastly, we found higher protein abundance of glucocorticoid receptors in light-exposed birds at death. Our results indicate that physiological and morphological traits vary in their ability to recover in response to constant light and warrants further investigation on the mechanisms driving stress resilience under a disrupted circadian rhythm.
{"title":"Zebra finches (Taeniopygia castanotis) display varying degrees of stress resilience and recovery in response to constant light.","authors":"Kevin Pham, Madeline Choi, KayLene Yamada, Haruka Wada","doi":"10.1016/j.ygcen.2024.114644","DOIUrl":"https://doi.org/10.1016/j.ygcen.2024.114644","url":null,"abstract":"<p><p>Traits vary in their ability to recover from stressors and is dependent on the magnitude, duration, or type of stressor. One such stressor is circadian rhythm disruption stemming from artificial light at night (ALAN). Circadian rhythm disruption may lead to long-term physiological consequences; however, the capacity in which individuals respond to recovery and display stress resilience is not known. Here, we exposed zebra finches (Taeniopygia castanotis) to constant light (24L:0D) or a regular light/dark cycle (14L:10D) for 23 days and a recovery period for 12 days. We measured body mass, corticosterone, and glucose levels before, during, and after constant light exposure and relative protein abundance of glucocorticoid receptors at death. In light-exposed birds, body mass significantly increased over time compared to birds under a regular light/dark cycle, but a 12-day recovery period reversed this increase. Baseline levels of circulating glucose decreased in light-exposed birds compared to controls, but significantly increased after the 12-day recovery period. In contrast, the glucose stress response did not show a similar recovery trend as body mass or baseline glucose in light-exposed birds. We did not detect any changes in baseline corticosterone or stress reactivity in both groups throughout the experiment. Lastly, we found higher protein abundance of glucocorticoid receptors in light-exposed birds at death. Our results indicate that physiological and morphological traits vary in their ability to recover in response to constant light and warrants further investigation on the mechanisms driving stress resilience under a disrupted circadian rhythm.</p>","PeriodicalId":12582,"journal":{"name":"General and comparative endocrinology","volume":" ","pages":"114644"},"PeriodicalIF":2.1,"publicationDate":"2024-11-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142727664","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-17DOI: 10.1016/j.ygcen.2024.114643
{"title":"Stress Axis: Molecular and Cellular Regulation of the HPI/HPA Axis.","authors":"","doi":"10.1016/j.ygcen.2024.114643","DOIUrl":"10.1016/j.ygcen.2024.114643","url":null,"abstract":"","PeriodicalId":12582,"journal":{"name":"General and comparative endocrinology","volume":" ","pages":"114643"},"PeriodicalIF":2.1,"publicationDate":"2024-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142647331","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-17DOI: 10.1016/j.ygcen.2024.114642
Sofía Proietto , Santiago Andrés Cortasa , Alejandro Raúl Schmidt , María Clara Corso , Pablo Ignacio Felipe Inserra , Noelia Paula Di Giorgio , Victoria Lux-Lantos , Alfredo Daniel Vitullo , Julia Halperin , Verónica Berta Dorfman
The plains vizcacha is a rodent that shows reactivation of the hypothalamic–pituitary–ovary (HPO) axis activity at mid-gestation. This process is enabled by the secretion of hypothalamic gonadotropin-releasing hormone (GnRH) at mid-gestation, followed by follicle-stimulating hormone (FSH) and luteinizing hormone (LH) secretion. However, a decrease in the pituitary GnRH receptor (GnRHR) expression is concomitantly determined. Moreover, an increment in the pituitary expression of estrogen receptor alpha (ERα) has been determined. This work aimed to study the impact of estradiol (E2) on GnRHR expression, the transcription factors early growth response protein-1 (Egr-1) and steroidogenic factor-1 (Sf-1), as well as on LH secretion. Three experimental approaches were performed: a physiological one with pregnant plains vizcachas, an in vivo approach with ovariectomized (OVX) animals treated with E2 (OVX + E2), and an ex vivo approach using pituitary glands exposed to a combination of GnRH and E2. Significant increased pituitary expression of Sf-1 and Egr-1 was determined at mid-gestation. Ovariectomy significantly increased adenohypophyseal expression levels of GnRHR, Egr-1, and Sf-1, as well as LH secretion. Then, OVX + E2 showed similar levels to SHAM. Adenohypophyses exposed to GnRH showed induced GnRHR, Egr-1, and Sf-1 expression, and LH secretion, while GnRH + E2 reverted these changes. The mid-gestation pituitary GnRHR decrease may result from the combination of increased E2 and GnRH secretion. Nevertheless, the increased expression of Egr-1 and Sf-1 at mid-gestation, together with LH release, suggests the tightly and complex regulatory system that takes place at mid-gestation, enabling a new progesterone surge that successfully carries the pregnancy to term.
New & Noteworthy
A significant increment of Sf-1 and Egr-1 at the pituitary of mid-gestating plains vizcachas was determined. Moreover, E2 reverted GnRHR, Egr-1, Sf-1, and LH increase in ovariectomized vizcachas’ pituitaries and ex vivo pituitaries exposed to GnRH. The decrease of the pituitary GnRHR at mid-gestation may result from the increased E2 and GnRH levels. A tightly and complex regulatory system may take place at mid-gestation enabling a new surge of progesterone that carries pregnancy to term.
{"title":"Estradiol affects the expression of essential molecular factors involved in luteinizing hormone secretion in the plains vizcacha","authors":"Sofía Proietto , Santiago Andrés Cortasa , Alejandro Raúl Schmidt , María Clara Corso , Pablo Ignacio Felipe Inserra , Noelia Paula Di Giorgio , Victoria Lux-Lantos , Alfredo Daniel Vitullo , Julia Halperin , Verónica Berta Dorfman","doi":"10.1016/j.ygcen.2024.114642","DOIUrl":"10.1016/j.ygcen.2024.114642","url":null,"abstract":"<div><div>The plains vizcacha is a rodent that shows reactivation of the hypothalamic–pituitary–ovary (HPO) axis activity at mid-gestation. This process is enabled by the secretion of hypothalamic gonadotropin-releasing hormone (GnRH) at mid-gestation, followed by follicle-stimulating hormone (FSH) and luteinizing hormone (LH) secretion. However, a decrease in the pituitary GnRH receptor (GnRHR) expression is concomitantly determined. Moreover, an increment in the pituitary expression of estrogen receptor alpha (ERα) has been determined. This work aimed to study the impact of estradiol (E2) on GnRHR expression, the transcription factors early growth response protein-1 (Egr-1) and steroidogenic factor-1 (Sf-1), as well as on LH secretion. Three experimental approaches were performed: a physiological one with pregnant plains vizcachas, an <em>in vivo</em> approach with ovariectomized (OVX) animals treated with E2 (OVX + E2), and an <em>ex vivo</em> approach using pituitary glands exposed to a combination of GnRH and E2. Significant increased pituitary expression of Sf-1 and Egr-1 was determined at mid-gestation. Ovariectomy significantly increased adenohypophyseal expression levels of GnRHR, Egr-1, and Sf-1, as well as LH secretion. Then, OVX + E2 showed similar levels to SHAM. Adenohypophyses exposed to GnRH showed induced GnRHR, Egr-1, and Sf-1 expression, and LH secretion, while GnRH + E2 reverted these changes. The mid-gestation pituitary GnRHR decrease may result from the combination of increased E2 and GnRH secretion. Nevertheless, the increased expression of Egr-1 and Sf-1 at mid-gestation, together with LH release, suggests the tightly and complex regulatory system that takes place at mid-gestation, enabling a new progesterone surge that successfully carries the pregnancy to term.</div></div><div><h3>New & Noteworthy</h3><div>A significant increment of Sf-1 and Egr-1 at the pituitary of mid-gestating plains vizcachas was determined. Moreover, E2 reverted GnRHR, Egr-1, Sf-1, and LH increase in ovariectomized vizcachas’ pituitaries and ex vivo pituitaries exposed to GnRH. The decrease of the pituitary GnRHR at mid-gestation may result from the increased E2 and GnRH levels. A tightly and complex regulatory system may take place at mid-gestation enabling a new surge of progesterone that carries pregnancy to term.</div></div>","PeriodicalId":12582,"journal":{"name":"General and comparative endocrinology","volume":"360 ","pages":"Article 114642"},"PeriodicalIF":2.1,"publicationDate":"2024-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142647326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-12DOI: 10.1016/j.ygcen.2024.114639
Ying Wang , Tianze Yang , Haolin Mo, Mingxing Yao, Qingchuan Song, Huixia Yu, Yuyou Du, Yang Li, Jiajia Yu, Lixin Wang
Melanocortin receptor-4 (MC4R) belongs to the G protein-coupled receptor family, characterized by a classical structure of seven transmembrane domains (7TMD). They play an important role in food intake and weight regulation. In the present study, we identified melanocortin-4-receptor-like (caMC4RL) mutants of goldfish from the Qian River in the Qin Ling region and characterized their functional properties, including the constitutive activities of the mutants, ligand-induced cAMP and ERK1/2 accumulation, and AMPK activation. The results show that six caMC4RL mutants were identified in goldfish from the Qian River in the Qin Ling region, and are located in the conserved position of the Cyprinidae MC4Rs. The mutations (E57K, P296S, and R302T/K) result in the loss of Gs signaling function. The mutations (P296 and R302T/K) exhibited biased signaling in response to ACTH stimulation in the MAPK/ERK pathway. In addition, the E57K mutant may play a role in weight regulation and could serve as molecular markers for molecular breeding. These data will provide fundamental information for functional studies of teleost GPCR mutants and MC4R isoforms.
{"title":"Identification and functional analysis of six melanocortin-4-receptor-like (MC4R-like) mutations in goldfish (Carassius auratus)","authors":"Ying Wang , Tianze Yang , Haolin Mo, Mingxing Yao, Qingchuan Song, Huixia Yu, Yuyou Du, Yang Li, Jiajia Yu, Lixin Wang","doi":"10.1016/j.ygcen.2024.114639","DOIUrl":"10.1016/j.ygcen.2024.114639","url":null,"abstract":"<div><div>Melanocortin receptor-4 (MC4R) belongs to the G protein-coupled receptor family, characterized by a classical structure of seven transmembrane domains (7TMD). They play an important role in food intake and weight regulation. In the present study, we identified <em>melanocortin-4-receptor-like</em> (<em>caMC4RL</em>) mutants of goldfish from the Qian River in the Qin Ling region and characterized their functional properties, including the constitutive activities of the mutants, ligand-induced cAMP and ERK1/2 accumulation, and AMPK activation. The results show that six <em>caMC4RL</em> mutants were identified in goldfish from the Qian River in the Qin Ling region, and are located in the conserved position of the Cyprinidae MC4Rs. The mutations (E57K, P296S, and R302T/K) result in the loss of Gs signaling function. The mutations (P296 and R302T/K) exhibited biased signaling in response to ACTH stimulation in the MAPK/ERK pathway. In addition, the E57K mutant may play a role in weight regulation and could serve as molecular markers for molecular breeding. These data will provide fundamental information for functional studies of teleost GPCR mutants and MC4R isoforms.</div></div>","PeriodicalId":12582,"journal":{"name":"General and comparative endocrinology","volume":"360 ","pages":"Article 114639"},"PeriodicalIF":2.1,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142618134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-12DOI: 10.1016/j.ygcen.2024.114640
Meenu Viswanath , M.C.Subhash Peter
In fish, as in other vertebrates, thyroid hormones (THs) act on many biological processes including growth and reproduction. Primary THs such as thyroxine (T4) and triiodothyronine (T3) are known for their direct action on osmoregulatory organs regulating ion osmotic homeostasis in many teleost fishes. However, it is unclear how these hormones interact with estradiol-17β (E2), an ovarian hormone that regulates the development of oocytes. We thus examined the short-term in vivo action of varied THs such as T4, T3 and T2, a potent TH metabolite diiodothyronine, on the expression pattern of receptors of THs and E2 in the ovarian wall of the hypothyroid climbing perch to identify the interactive pattern of TH/E2 receptor system and the molecular dynamics of Na+/K+−ATPase (NKA) subunits in the ovarian wall that provides structural and functional support to ovary. We found differential pattern of transcript abundance of NKA subunits isoforms such as nkaα1a, nkaα1b, nkaα1c atp1b1, atp1b2 and fxyd3, fxyd5, fxyd6, TH receptor isoforms (tr<, trβ, tr<B), deiodinases (deo1, deo2 and deo3) and TH transporter (mct8, mct10) and E2 receptor (er<, erβ, gper), aromatases (cyp19a1a, cyp19a1b) and steroidogenic enzymes (17β-hsd, sf-1, star) in ovarian wall in response to T4, T3 and T2 administration in MMI-treated fish. The transcript abundance pattern provides evidence for a direct role of THs in the ovarian wall of hypothyroid fish. It confirms a targeted interaction of THs with ER/cyp19a1 and NKA systems. Pattern analysis further revealed that T4 could produce maximum activation of the TR/Deo/Mct and ER/cyp19a1 systems among three iodinated tyrosines, bringing synergistic interaction between the TH and E2 systems in the ovarian wall. This novel evidence of direct interaction between the TH/E2 system and the NKA system in the ovarian wall further testifies to an ion osmotic role for THs in this ovarian structure.
{"title":"Thyroid hormones activate TH/E2 receptor/regulator system and drive Na+/K+-ATPase in the ovarian wall of hypothyroid air-breathing fish (Anabas testudineus Bloch)","authors":"Meenu Viswanath , M.C.Subhash Peter","doi":"10.1016/j.ygcen.2024.114640","DOIUrl":"10.1016/j.ygcen.2024.114640","url":null,"abstract":"<div><div>In fish, as in other vertebrates, thyroid hormones (THs) act on many biological processes including growth and reproduction. Primary THs such as thyroxine (T<sub>4</sub>) and triiodothyronine (T<sub>3</sub>) are known for their direct action on osmoregulatory organs regulating ion osmotic homeostasis in many teleost fishes. However, it is unclear how these hormones interact with estradiol-17β (E<sub>2</sub>), an ovarian hormone that regulates the development of oocytes. We thus examined the short-term <em>in vivo</em> action of varied THs such as T<sub>4,</sub> T<sub>3</sub> and T<sub>2</sub>, a potent TH metabolite diiodothyronine, on the expression pattern of receptors of THs and E<sub>2</sub> in the ovarian wall of the hypothyroid climbing perch to identify the interactive pattern of TH/E<sub>2</sub> receptor system and the molecular dynamics of Na<sup>+</sup>/K<sup>+</sup>−ATPase (NKA) subunits in the ovarian wall that provides structural and functional support to ovary. We found differential pattern of transcript abundance of NKA subunits isoforms such as <em>nkaα1a, nka</em>α<em>1b, nka</em>α<em>1c atp1b1, atp1b2</em> and <em>fxyd3</em>, <em>fxyd5, fxyd6,</em> TH receptor isoforms (<em>tr<, trβ, tr<B</em>), deiodinases (<em>deo1, deo2</em> and <em>deo3</em>) and TH transporter (<em>mct8, mct10</em>) and E<sub>2</sub> receptor (<em>er<, erβ, gper</em>), aromatases (<em>cyp19a1a, cyp19a1b</em>) and steroidogenic enzymes (<em>17β-hsd, sf-1, star</em>) in ovarian wall in response to T<sub>4</sub>, T<sub>3</sub> and T<sub>2</sub> administration in MMI-treated fish. The transcript abundance pattern provides evidence for a direct role of THs in the ovarian wall of hypothyroid fish. It confirms a targeted interaction of THs with ER/cyp19a1 and NKA systems. Pattern analysis further revealed that T<sub>4</sub> could produce maximum activation of the TR/Deo/Mct and ER/cyp19a1 systems among three iodinated tyrosines, bringing synergistic interaction between the TH and E<sub>2</sub> systems in the ovarian wall. This novel evidence of direct interaction between the TH/E<sub>2</sub> system and the NKA system in the ovarian wall further testifies to an ion osmotic role for THs in this ovarian structure.</div></div>","PeriodicalId":12582,"journal":{"name":"General and comparative endocrinology","volume":"360 ","pages":"Article 114640"},"PeriodicalIF":2.1,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142618132","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-12DOI: 10.1016/j.ygcen.2024.114641
Timothy S. Breton , Maria Eduarda Oliveira , Truly Chillemi , William Harriman , Joanna Korasadowicz , Eme Saverese , Emma Bourget , Casey A. Murray , Christopher J. Martyniuk , Matthew A. DiMaggio
Super-conserved Receptors Expressed in Brain (SREB) are a highly conserved family of orphan G protein-coupled receptors that consist of three members in most vertebrates: SREB1 (GPR27), SREB2 (GPR85), and SREB3 (GPR173). Each receptor is associated with diverse physiological processes and expressed in both ovaries and testes, but reproductive functions are only beginning to be understood. In addition, some fishes gained a novel fourth gene, SREB3B, which may have unique functions. The purpose of this study was to conduct a spatial and quantitative analysis of SREBs in the gonads of pufferfish (Dichotomyctere nigroviridis), which expresses all four genes. Multiplex RNAscope and absolute qPCR were used to assess gene expression patterns in both ovaries and testes. Expression was detected in early ovaries and dominated by sreb1 (approximately 2500 copies/ng RNA vs. 300 or less for others), with notable expression of all receptors in primary oocytes, granulosa cells, and small numbers of extra-follicular cells. Within primary oocytes, sreb1 and sreb3b exhibited diffuse patterns that may indicate early functions, while sreb2 and sreb3a were granular and may reflect stored mRNA. Early testicular development was dominated by sreb1 and sreb2 (∼5000 copies/ng RNA) in spermatogonia. These patterns were somewhat reduced in late testes (∼1000–2600 copies/ng RNA), but sreb3b exhibited a novel spatial pattern (∼380 copies/ng RNA) within spermatogenic cysts. These results highlight diverse roles for the SREB family, and sreb3b is hypothesized to have unique roles in fish reproduction.
{"title":"Spatial and quantitative gene expression analysis of SREB receptors in the gonads of green-spotted pufferfish (Dichotomyctere nigroviridis)","authors":"Timothy S. Breton , Maria Eduarda Oliveira , Truly Chillemi , William Harriman , Joanna Korasadowicz , Eme Saverese , Emma Bourget , Casey A. Murray , Christopher J. Martyniuk , Matthew A. DiMaggio","doi":"10.1016/j.ygcen.2024.114641","DOIUrl":"10.1016/j.ygcen.2024.114641","url":null,"abstract":"<div><div>Super-conserved Receptors Expressed in Brain (SREB) are a highly conserved family of orphan G protein-coupled receptors that consist of three members in most vertebrates: SREB1 (GPR27), SREB2 (GPR85), and SREB3 (GPR173). Each receptor is associated with diverse physiological processes and expressed in both ovaries and testes, but reproductive functions are only beginning to be understood. In addition, some fishes gained a novel fourth gene, SREB3B, which may have unique functions. The purpose of this study was to conduct a spatial and quantitative analysis of SREBs in the gonads of pufferfish (<em>Dichotomyctere nigroviridis</em>), which expresses all four genes. Multiplex RNAscope and absolute qPCR were used to assess gene expression patterns in both ovaries and testes. Expression was detected in early ovaries and dominated by <em>sreb1</em> (approximately 2500 copies/ng RNA vs. 300 or less for others), with notable expression of all receptors in primary oocytes, granulosa cells, and small numbers of extra-follicular cells. Within primary oocytes, <em>sreb1</em> and <em>sreb3b</em> exhibited diffuse patterns that may indicate early functions, while <em>sreb2</em> and <em>sreb3a</em> were granular and may reflect stored mRNA. Early testicular development was dominated by <em>sreb1</em> and <em>sreb2</em> (∼5000 copies/ng RNA) in spermatogonia. These patterns were somewhat reduced in late testes (∼1000–2600 copies/ng RNA), but <em>sreb3b</em> exhibited a novel spatial pattern (∼380 copies/ng RNA) within spermatogenic cysts. These results highlight diverse roles for the SREB family, and <em>sreb3b</em> is hypothesized to have unique roles in fish reproduction.</div></div>","PeriodicalId":12582,"journal":{"name":"General and comparative endocrinology","volume":"360 ","pages":"Article 114641"},"PeriodicalIF":2.1,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142618129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-18DOI: 10.1016/j.ygcen.2024.114629
Bin Wang , Zhenfang Tian , Zhihua Yu , Aijun Cui , Yan Jiang , Hai Huang , Yongjiang Xu
Spexin (SPX1) is a novel neuropeptide composed of 14 amino acids and well conserved across vertebrates, and it has been implicated in various physiological functions via galanin receptor 2 (GALR2) and GALR3. However, the detailed signaling pathways mediating its actions in target cells are still largely unknown. Accordingly, we addressed this issue in the present study using yellowtail kingfish as a model. SPX1 significantly increased CRE-luc activity in COS-7 cells expressing its cognate receptors GALR2a and GALR2b, and this stimulatory effect was attenuated by two inhibitors of the PKA pathway. Similarly, an evident induction of SRE-luc activity was observed when COS-7 cells transfected with GALR1b, GALR2a, GALR2b, GALR type 1, or GALR type 2 were challenged with SPX1, and two blockers of the PKC pathway suppressed this stimulatory action. Moreover, SPX1 markedly elevated NFAT-RE-luc activity in COS-7 cells expressing GALR1a, GALR2a, or GALR2b, and this promotion was inhibited by two antagonists of the Ca2+ route. Overall, our results have revealed that activation of six yellowtail kingfish galanin receptors by the SPX1 peptide may occur with different downstream signaling events, which could account for its pleotropic functions.
{"title":"Differential activation of six galanin receptors by the spexin peptide in yellowtail kingfish (Seriola lalandi)","authors":"Bin Wang , Zhenfang Tian , Zhihua Yu , Aijun Cui , Yan Jiang , Hai Huang , Yongjiang Xu","doi":"10.1016/j.ygcen.2024.114629","DOIUrl":"10.1016/j.ygcen.2024.114629","url":null,"abstract":"<div><div>Spexin (SPX1) is a novel neuropeptide composed of 14 amino acids and well conserved across vertebrates, and it has been implicated in various physiological functions via galanin receptor 2 (GALR2) and GALR3. However, the detailed signaling pathways mediating its actions in target cells are still largely unknown. Accordingly, we addressed this issue in the present study using yellowtail kingfish as a model. SPX1 significantly increased CRE-luc activity in COS-7 cells expressing its cognate receptors GALR2a and GALR2b, and this stimulatory effect was attenuated by two inhibitors of the PKA pathway. Similarly, an evident induction of SRE-luc activity was observed when COS-7 cells transfected with GALR1b, GALR2a, GALR2b, GALR type 1, or GALR type 2 were challenged with SPX1, and two blockers of the PKC pathway suppressed this stimulatory action. Moreover, SPX1 markedly elevated NFAT-RE-luc activity in COS-7 cells expressing GALR1a, GALR2a, or GALR2b, and this promotion was inhibited by two antagonists of the Ca<sup>2+</sup> route. Overall, our results have revealed that activation of six yellowtail kingfish galanin receptors by the SPX1 peptide may occur with different downstream signaling events, which could account for its pleotropic functions.</div></div>","PeriodicalId":12582,"journal":{"name":"General and comparative endocrinology","volume":"359 ","pages":"Article 114629"},"PeriodicalIF":2.1,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142462751","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Fat mass and obesity associated gene (FTO) has been strongly associated with obesity, and it is functionally linked to the homeobox transcription factor iriquois-3 (IRX3). In mammals, FTO and IRX3 are involved in the regulation of food intake and metabolism. This study aimed to determine whether FTO and IRX3 are affected by feeding and food unavailability. FTO and IRX3 mRNA and protein were found widely distributed in all tissues examined, including the brain, muscle, gut, and liver. Postprandial increase in the abundance of FTO and IRX3 mRNAs was observed in metabolic tissues of both male and female zebrafish at 1 h post-feeding. Meanwhile, their expression in the brain and gut decreased at 3 h post-feeding, reaching preprandial levels. Additionally, FTO and IRX3 mRNA abundance in examined tissues increased after 7 days of food deprivation, but substantially decreased after refeeding for 24 h. In summary, we report that both FTO and IRX3 are meal-sensitive genes in zebrafish. The fasting-induced increase suggests a possible appetite regulatory role for FTO and IRX3 in zebrafish. These findings highlight the importance of FTO and IRX3 in appetite and metabolic regulation in zebrafish.
{"title":"Fat mass and obesity associated gene and homeobox transcription factor iriquois-3 mRNA profiles in the metabolic tissues of zebrafish are modulated by feeding and food deprivation","authors":"Katayoon Karimzadeh , Chinelo Uju , Asgar Zahmatkesh , Suraj Unniappan","doi":"10.1016/j.ygcen.2024.114621","DOIUrl":"10.1016/j.ygcen.2024.114621","url":null,"abstract":"<div><div>Fat mass and obesity associated gene (FTO) has been strongly associated with obesity, and it is functionally linked to the homeobox transcription factor iriquois-3 (IRX3). In mammals, FTO and IRX3 are involved in the regulation of food intake and metabolism. This study aimed to determine whether FTO and IRX3<!--> <!-->are affected by feeding and food unavailability. FTO and IRX3 mRNA and protein were found widely distributed in all tissues examined, including the brain, muscle, gut, and liver. Postprandial increase in the abundance of FTO and IRX3 mRNAs was observed in metabolic tissues of both male and female zebrafish at 1 h post-feeding. Meanwhile, their expression in the brain and gut decreased at 3 h post-feeding, reaching preprandial levels. Additionally, FTO and IRX3 mRNA abundance in examined tissues increased after 7 days of food deprivation, but substantially decreased after refeeding for 24 h. In summary, we report that both FTO and IRX3 are meal-sensitive genes in zebrafish. The fasting-induced increase suggests a possible appetite regulatory role for FTO and IRX3 in zebrafish. These findings highlight the importance of FTO and IRX3 in appetite and metabolic regulation in zebrafish.</div></div>","PeriodicalId":12582,"journal":{"name":"General and comparative endocrinology","volume":"360 ","pages":"Article 114621"},"PeriodicalIF":2.1,"publicationDate":"2024-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142462752","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-14DOI: 10.1016/j.ygcen.2024.114628
Cui Liu , Longsheng Zhang , You Xia , Keqi Li , Jikui Wu , Junling Zhang
Rbpms2, an RNA-binding protein with multiple splicing (Rbpms), can interact with RNAs to involve oocyte development, thereby influencing female sex differentiation in vertebrates. Here, two splicing variants of the Rbpms2 gene from Japanese flounder (Paralichthys olivaceus) were identified, namely Rbpms2.1 and Rbpms2.2. The two variants exhibited 98.22 % amino acid homology, both featuring an RNA recognition motif (RRM) domain spanning positions 98–170 amino acids. They were relatively conserved throughout phylogenetic evolution. Differently, the C-terminal region of the Rbpms2.1 contains five additional sequential amino acids (–VRDQP–) compared to Rbpms2.2. The real-time qPCR results demonstrated that Rbpms2.1 and Rbpms2.2 had relatively abundant expression in the gonads of adult Japanese flounder, with higher expression levels in the ovary compared to the testis (P < 0.05). In situ hybridization results showed strong positive expression of Rbpms2 mRNA in oocytes at stages I-III during the V stage of ovarian development. In the testis at stage IV, the expression of Rbpms2 mRNA was mainly concentrated on primary spermatocytes. Importantly, Rbpms2 binding sites were found in the 3′UTR, 5′UTR, and ORF regions of the sex-related genes including dmrt1, sox9, amh, foxl2, and wnt4. siRNA interference and overexpression analysis of Rbpms2.1 and Rbpms2.2 in primary cells of the ovary and testis showed that Rbpms2 can repress the expression of male-related genes (dmrt1, sox9, and amh) and significantly promote the expression of female-related genes (foxl2 and wnt4). Our results revealed that Rbpms2 may play a critical role by targeting the sex-related genes in the gonad development of Japanese flounder.
{"title":"Identification, expression, and function analysis of Rbpms2 splicing variants in Japanese flounder gonad","authors":"Cui Liu , Longsheng Zhang , You Xia , Keqi Li , Jikui Wu , Junling Zhang","doi":"10.1016/j.ygcen.2024.114628","DOIUrl":"10.1016/j.ygcen.2024.114628","url":null,"abstract":"<div><div>Rbpms2<em>,</em> an RNA-binding protein with multiple splicing (<em>Rbpms</em>), can interact with RNAs to involve oocyte development, thereby influencing female sex differentiation in vertebrates. Here, two splicing variants of the <em>Rbpms2</em> gene from Japanese flounder (<em>Paralichthys olivaceus</em>) were identified, namely <em>Rbpms2.1</em> and <em>Rbpms2.2</em>. The two variants exhibited 98.22 % amino acid homology, both featuring an RNA recognition motif (RRM) domain spanning positions 98–170 amino acids. They were relatively conserved throughout phylogenetic evolution. Differently, the C-terminal region of the <em>Rbpms2.1</em> contains five additional sequential amino acids (–VRDQP–) compared to <em>Rbpms2.2</em>. The real-time qPCR results demonstrated that <em>Rbpms2.1</em> and <em>Rbpms2.2</em> had relatively abundant expression in the gonads of adult Japanese flounder, with higher expression levels in the ovary compared to the testis (<em>P</em> < 0.05). <em>In situ</em> hybridization results showed strong positive expression of <em>Rbpms2</em> mRNA in oocytes at stages I-III during the V stage of ovarian development. In the testis at<!--> <!-->stage IV, the expression of <em>Rbpms2</em> mRNA was mainly concentrated on primary spermatocytes. Importantly, <em>Rbpms2</em> binding sites were found in the 3′UTR, 5′UTR, and ORF regions of the sex-related genes including <em>dmrt1</em>, <em>sox9</em>, <em>amh</em>, <em>foxl2</em>, and <em>wnt4</em>. siRNA interference and overexpression analysis of <em>Rbpms2.1</em> and <em>Rbpms2.2</em> in primary cells of the ovary and testis showed that <em>Rbpms2</em> can repress the expression of male-related genes (<em>dmrt1</em>, <em>sox9</em>, and <em>amh</em>) and significantly promote the expression of female-related genes (<em>foxl2</em> and <em>wnt4</em>). Our results revealed that <em>Rbpms2</em> may play a critical role by targeting the sex-related genes in the gonad development of Japanese flounder.</div></div>","PeriodicalId":12582,"journal":{"name":"General and comparative endocrinology","volume":"359 ","pages":"Article 114628"},"PeriodicalIF":2.1,"publicationDate":"2024-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142444725","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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