GABAB receptor antagonism from birth to weaning permanently modifies Kiss1 expression in hypothalamus and gonads in mice

2021 ◽  
Author(s):  
Marianne Bizzozzero Hiriart ◽  
Noelia P. Di Giorgio ◽  
Carlos Libertun ◽  
Victoria A.R. Lux-Lantos
Keyword(s):  
Arcuate Nucleus ◽  
Gabab Receptor ◽  
Seminiferous Tubules ◽  
Testis Weight ◽  
Hpg Axis ◽  
Estrous Cycles ◽  
Amino Butyric Acid ◽  
Periventricular Nucleus ◽  
Puberty Onset ◽  
Ovulatory Follicles

Introduction: The kisspeptin gene Kiss1 is expressed in two hypothalamic areas: anteroventral periventricular nucleus/periventricular nucleus (AVPV/PeN) and arcuate nucleus (ARC), and also in gonads. Evidences suggest that gamma-amino butyric acid B receptors (GABAB) signaling can regulate Kiss1 expression. Here we inhibited GABAB signaling from PND2-PND21 and evaluated the hypothalamic-pituitary-gonadal (HPG) axis. Methods: BALB/c mice were treated on postnatal days 2-21 (PND2-PND21) with CGP55845 (GABAB antagonist), and evaluated in PND21 and adulthood: gene expression (qPCR) in hypothalamus and gonads, hormones by radioimmunoassay, gonad histochemistry (H&E), puberty onset, estrous cycles. Results: At PND21, CGP inhibited Kiss1 and Tac2 and increased Pdyn and Gabbr1 in the ARC of both sexes and decreased Th only in female AVPV/PeN. Serum follicle-stimulating hormone (FSH) and testis weight decreased in CGP-males and puberty onset was delayed. In adults, Kiss1, Tac2, Pdyn, Pgr, Cyp19a1, Gad1 were downregulated, while Gabbr1 was upregulated in the ARC of both sexes. In the AVPV/PeN, Kiss1, Th, Cyp19a1 and Pgr decreased while Gad1 increased in CGP-females, whereas Cyp19a1 increased in CGP-males. Serum FSH increased in CGP-males while prolactin increased in CGP-females. Testosterone and progesterone increased in ovaries from CGP-females, in which Kiss1, Cyp19a1 and Esr1 were downregulated while Hsd3b2 was upregulated, together with increased atretic and decreased ovulatory follicles. Testes from CGP-males showed decreased progesterone, increased Gabbr1, Kiss1, Kiss1r, Esr2 and decreased Cyp19a1 and clear signs of seminiferous tubules atrophy. Conclusion: These results demonstrate that appropriate GABAB signaling during this critical prepubertal period is necessary for the normal development of the HPG axis.

scholarly journals GABAergic input through GABAB receptors is necessary during a perinatal window to shape gene expression of factors critical to reproduction such as Kiss1

2020 ◽  
Vol 318 (6) ◽  
pp. E901-E919 ◽  
Author(s):  
Marianne Bizzozzero-Hiriart ◽  
Noelia P. Di Giorgio ◽  
Carlos Libertun ◽  
Victoria Lux-Lantos
Keyword(s):  
Gene Expression ◽  
Postnatal Period ◽  
Gabab Receptors ◽  
Acid Decarboxylase ◽  
Estrous Cycles ◽  
Primordial Follicles ◽  
Adult Mice ◽  
Periventricular Nucleus ◽  
Puberty Onset ◽  
Blood Hormones

Lack of GABAB receptors in GABAB1 knockout mice decreases neonatal ARC kisspeptin 1 ( Kiss1) expression in the arcuate nucleus of the hypothalamus (ARC) in females, which show impaired reproduction as adults. Our aim was to selectively impair GABAB signaling during a short postnatal period to evaluate its impact on the reproductive system. Neonatal male and female mice were injected with the GABAB antagonist CGP 55845 (CGP, 1 mg/kg body wt sc) or saline from postnatal day 2 ( PND2) to PND6, three times per day (8 AM, 1 PM, and 6 PM). One group was killed on PND6 for collection of blood samples (hormones by radioimmunoassay), brains for gene expression in the anteroventral periventricular nucleus-periventricular nucleus continuum (AVPV/PeN), and ARC micropunches [quantitative PCR (qPCR)] and gonads for qPCR, hormone contents, and histology. A second group of mice was injected with CGP (1 mg/kg body wt sc) or saline from PND2 to PND6, three times per day (8 AM, 1 PM, and 6 PM), and left to grow to adulthood. We measured body weight during development and parameters of sexual differentiation, puberty onset, and estrous cycles. Adult mice were killed, and trunk blood (hormones), brains for qPCR, and gonads for qPCR and hormone contents were obtained. Our most important findings on PND6 include the CGP-induced decrease in ARC Kiss1 and increase in neurokinin B ( Tac2) in both sexes; the decrease in AVPV/PeN tyrosine hydroxylase ( Th) only in females; the increase in gonad estradiol content in both sexes; and the increase in primordial follicles and decrease in primary and secondary follicles. Neonatally CGP-treated adults showed decreased ARC Kiss1 and ARC gonadotropin-releasing hormone ( Gnrh1) and increased ARC glutamic acid decarboxylase 67 ( Gad1) only in males; increased ARC GABAB receptor subunit 1 ( Gabbr1) in both sexes; and decreased AVPV/PeN Th only in females. We demonstrate that ARC Kiss1 expression is chronically downregulated in males and that the normal sex difference in AVPV/PeN Th expression is abolished. In conclusion, neonatal GABAergic input through GABAB receptors shapes gene expression of factors critical to reproduction.

scholarly journals SUN-244 Role of Activin, Follistatin, and Inhibin in the Regulation of KISS-1 Gene Expression in Hypothalamic Cell Models

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Aki Oride ◽  
Haruhiko Kanasaki ◽  
Zolzaya Tumurgan ◽  
Tuvshintugs Tumurbaatar ◽  
Satoru Kyo
Keyword(s):  
Gene Expression ◽  
Arcuate Nucleus ◽  
Cell Model ◽  
Gonadotropin Releasing Hormone ◽  
The Other ◽  
Cell Models ◽  
Inhibin A ◽  
Hpg Axis ◽  
Periventricular Nucleus

Abstract Kisspeptin (encoded by the Kiss-1 gene) in the arcuate nucleus (ARC) of the hypothalamus governs the hypothalamic-pituitary-gonadal (HPG) axis by regulating pulsatile release of gonadotropin-releasing hormone (GnRH). Meanwhile, kisspeptin in the anteroventral periventricular nucleus (AVPV) region has been implicated in estradiol (E2)-induced GnRH surges. Kiss-1-expressing cell model mHypoA-55 exhibits characteristics of Kiss-1 neurons in the ARC region. On the other hand, Kiss-1 expressing mHypoA-50 cells originate from the AVPV region. In the mHypoA-55 ARC cells, activin significantly increased Kiss-1 gene expression. Follistatin alone reduced Kiss-1 expression within these cells. Interestingly, activin-induced Kiss-1 gene expression was completely abolished by follistatin. Inhibin A, but not inhibin B reduced Kiss-1 expression. Activin-increased Kiss-1 expression was also abolished by inhibin A. Pretreatment of the cells with follistatin or inhibin A significantly inhibited kisspeptin- or GnRH-induced Kiss-1 gene expression in mHypoA-55 cells. In contrast, in the mHypoA-50 AVPV cell model, activin, follistatin, and inhibin A did not modulate Kiss-1 gene expression. The subunits that compose activin and inhibin, as well as follistatin were expressed in both mHypoA-55 and mHypoA-50 cells. Expression of inhibin βA and βB subunits and follistatin was much higher in mHypoA-55 ARC cells. Furthermore, we found that expression of the inhibin αsubunit and follistatin genes was modulated in the presence of E2 in mHypoA-55 ARC cells. The results of this study suggest that activin, follistatin, and inhibin A within the ARC region participate in the regulation of the HPG axis under the influence of E2.

Role of activin, follistatin, and inhibin in the regulation of Kiss-1 gene expression in hypothalamic cell models†

2019 ◽  
Vol 101 (2) ◽  
pp. 405-415 ◽  
Author(s):  
Zolzaya Tumurgan ◽  
Haruhiko Kanasaki ◽  
Tuvshintugs Tumurbaatar ◽  
Aki Oride ◽  
Hiroe Okada ◽  
...  
Keyword(s):  
Gene Expression ◽  
Arcuate Nucleus ◽  
Cell Model ◽  
Gonadotropin Releasing Hormone ◽  
Cell Models ◽  
Inhibin A ◽  
Hpg Axis ◽  
Α Subunit ◽  
Periventricular Nucleus

Abstract Kisspeptin (encoded by the Kiss-1 gene) in the arcuate nucleus (ARC) of the hypothalamus governs the hypothalamic-pituitary-gonadal (HPG) axis by regulating pulsatile release of gonadotropin-releasing hormone (GnRH). Meanwhile, kisspeptin in the anteroventral periventricular nucleus (AVPV) region has been implicated in estradiol (E2)-induced GnRH surges. Kiss-1–expressing cell model mHypoA-55 exhibits characteristics of Kiss-1 neurons in the ARC region. On the other hand, Kiss-1 expressing mHypoA-50 cells originate from the AVPV region. In the mHypoA-55 ARC cells, activin significantly increased Kiss-1 gene expression. Follistatin alone reduced Kiss-1 expression within these cells. Interestingly, activin-induced Kiss-1 gene expression was completely abolished by follistatin. Inhibin A, but not inhibin B reduced Kiss-1 expression. Activin-increased Kiss-1 expression was also abolished by inhibin A. Pretreatment of the cells with follistatin or inhibin A significantly inhibited kisspeptin- or GnRH-induced Kiss-1 gene expression in mHypoA-55 cells. In contrast, in the mHypoA-50 AVPV cell model, activin, follistatin, and inhibin A did not modulate Kiss-1 gene expression. The subunits that compose activin and inhibin, as well as follistatin were expressed in both mHypoA-55 and mHypoA-50 cells. Expression of inhibin βA and βB subunits and follistatin was much higher in mHypoA-55 ARC cells. Furthermore, we found that expression of the inhibin α subunit and follistatin genes was modulated in the presence of E2 in mHypoA-55 ARC cells. The results of this study suggest that activin, follistatin, and inhibin A within the ARC region participate in the regulation of the HPG axis under the influence of E2.

scholarly journals Grid1 Regulates the Onset of Puberty in Female Rats

2020 ◽  
Author(s):  
Jing Ye ◽  
Ping Qin ◽  
Hailing Li ◽  
Tiezhu Kang ◽  
Wenyu Si ◽  
...  
Keyword(s):  
Arcuate Nucleus ◽  
Female Rats ◽  
Mrna Levels ◽  
Vaginal Opening ◽  
Ionotropic Receptor ◽  
Delta Type ◽  
Periventricular Nucleus ◽  
Old Rats ◽  
Puberty Onset

Abstract The present study aimed to investigate whether Grid1, encoding the glutamate ionotropic receptor delta type subunit 1(GluD1), influences the onset of puberty in female rats. First, we detected the expression of Grid1 mRNA and its protein in the hypothalamus from infancy to puberty. Second, we evaluated the suppression of Grid1 expression by Lentivirus-Grid1 (LV-Grid1) in primary hypothalamus cells through measuring the expression level of Grid1. Finally, LV-Grid1 was intracerebroventricular injected (ICV) into 21-day-old rats and to investigate the effect of Grid1 suppression on puberty onset in vivo. Results showed that GluD1 immunoreactivity could be detected in the arcuate nucleus (ARC), paraventricular nucleus (PVN), and periventricular nucleus (PeN). Grid1 mRNA levels were the lowest at prepuberty. Treatment of hypothalamic neurons with LV-Grid1 decreased the mRNA expression levels of Grid1 and Rfrp-3 (encoding RFamide-related peptide 3, RFRP 3), but increased that of Gnrh (encoding gonadotropin-releasing hormone, GnRH). After 7 days of ICV LV-Grid1 into rats, the Grid1 mRNA was significantly reduced (by 46%), Gnrh mRNA expression was significantly increased, but Rfrp-3 mRNA levels were decreased. The time of rat vaginal opening (VO) was earlier in the LV-Grid1 group; the concentrations of luteinizing hormone (LH), estradiol (E2), and progesterone (P4) in serum were significantly increased; and the ovaries were significantly larger. Our study revealed that Grid1 affects the onset of puberty by regulating the level of GnRH and RFRP3.

A powerful GABAB receptor-mediated inhibition of GABAergic neurons in arcuate nucleus

Neuroreport ◽  
1999 ◽  
Vol 10 (12) ◽  
pp. 2681-2687 ◽  
Author(s):  
Edward J. Wagner ◽  
Martha A. Bosch ◽  
Martin J. Kelly ◽  
Oline K. Rønnekleiv
Keyword(s):  
Arcuate Nucleus ◽  
Gabab Receptor ◽  
Gabaergic Neurons

scholarly journals Reproductive cycle of male wall lizard, Podarcis vaucheri (Reptilia: Sauria: Lacertidae), in Djurdjura, Northern Algeria

2017 ◽  
Author(s):  
Rabah Mamou ◽  
Elara Moudilou ◽  
Jean-Marie Exbrayat ◽  
Mansour Amroun
Keyword(s):  
Sexual Activity ◽  
Reproductive Cycle ◽  
Mixed Type ◽  
Progressive Increase ◽  
Early Summer ◽  
Seminiferous Tubules ◽  
Testis Weight ◽  
Field Observations ◽  
Testicular Weight ◽  
Resting Period

The reproductive cycle of male wall lizard Podarcis vaucheri (Boulenger, 1905) living in Djurdjura mountain, was analyzed from April till October 2014. Its seasonal mixed-type cycle was characterized by three phases: sexual activity, quiescence and recrudescence. After the emergence of lizard from hibernation an intense sexual activity was observed in spring (April, May, and June) at which the testicular weight and seminiferous tubules diameter reached their maximal values. At this period, spermiogenesis was also maximal and the spermiation allowed the release of a large quantity of spermatozoa in both the seminiferous tubules and epididymis lumen. In July, a short sexual resting period followed, during which the testes mass and seminiferous tubules diameter decreased. Spermatogonia and some primary spermatocytes were observed against the wall of seminiferous tubules. The recrudescence period (August, September and October) was characterized by a progressive increase of the testis weight and seminiferous tubules diameter. Spermatocytogenesis began in August, and spermiogenesis occurred in September and October, before the diapause period. But the presence of spermatozoids was not signaled in the epididymis lumen. Field observations and microscopic evidence of testis showed that both sexes reproduced synchronously during spring and early summer.

520. RELATIONSHIP BETWEEN THE OVARIAN STATUS ON THE INITIATION DAY OF SUPEROVULATION AND THE OVULATORY FOLLICLES AT ESTRUS IN SHALL IRANIAN EWES

2009 ◽  
Vol 21 (9) ◽  
pp. 119
Author(s):  
H. Kermani Moakhar ◽  
H. Kohram ◽  
A. Zare Shahne
Keyword(s):  
Estrous Cycles ◽  
Ovarian Status ◽  
Starting Point ◽  
Ovulatory Follicles

The objective of this experiment was to determine the influence of the ovarian status on the initiation day of superovulation (day 14) on superovulatory response in terms of the number of ovulatory follicles (F, ≥4mm) at estrus (day 16). Estrous cycles of 50 Shall Iranian ewes were synchronized by CIDR. The day of CIDR insertion was considered as the starting point of the experiment (day 0). At the time of CIDR removal (day 14), 650 IU of eCG was administered to ewes. Ultrasonography was performed on days 14 and 16. Data were analyzed by means of the GLM procedure of the SAS. In these cases, the number of follicles were determined in each of the three studied classes (<2, 2 to 4, and ≥4 mm) on day 14. Subsequently, each of these classes was divided into two subgroups (Low, High) based on the number of follicles observed on day 14 (Table 1). The number of large follicles at estrus in each of the three studied classes were (3.96±0.28 vs. 4.78±0.26, p<0.05; 4.31±0.35 vs. 4.41±0.24, p>0.05; and 4.50±0.23 vs. 4.07±0.37, p>0.05) respectively. The number of medium and large follicles on day 14 did not affect the superovulatory response. However, the high number of small follicles on the initiation day of superovulation increased the number of ovulatory follicles at estrus. Since, 23 out of 50 ewes had a low number of small follicles on day 14 (Table 1), which may suggest that an injection of FSH 1-2 days before initiation of eCG treatment enhance the superovulatory response.

Relationship between testicular morphology and sperm production following ischaemia in the ram

1995 ◽  
Vol 7 (1) ◽  
pp. 119 ◽  
Author(s):  
CM Markey ◽  
AM Jequier ◽  
GT Meyer ◽  
GB Martin
Keyword(s):  
Morphological Changes ◽  
Sperm Concentration ◽  
Testicular Biopsy ◽  
Seminiferous Tubules ◽  
Testis Weight ◽  
Infertile Men ◽  
Testicular Morphology ◽  
And Function ◽  
The Relationship ◽  
Spermatogenic Epithelium

Arteriosclerosis was induced in the internal spermatic artery of rams to determine if this condition is implicated in the aetiology of testicular pathology which causes male infertility. Data were collected on sperm concentration and motility for 56 days following surgery to provide an index of testicular function. Testes were then weighed and a testicular biopsy score count was performed on histological sections to assess spermatogenic potential of seminiferous tubules. Vascular disturbance caused focal damage of the seminiferous epithelium, similar to that seen among infertile men, and a reduction in ejaculate volume, sperm concentration and sperm motility. Sperm concentration decreased following ischaemia yet was maintained to some degree by a germ-cell depleted spermatogenic epithelium. Normal testicular morphology was maintained above a testis weight of about 120 g (for an individual testis), but below this threshold spermatogenesis was severely impaired. In conclusion, these data have provided information on the relationship between testicular morphology and function following ischaemia in the ram. Furthermore, the morphological changes induced in the testis were similar to those seen among infertile men and, by their focal nature, could explain the distinction between oligozoospermia and azoospermia in men exhibiting spermatogenic arrest.

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