Step by Step about Germ Cells Development in Canine

Souza, Aline Fernanda de; Pieri, Naira Caroline Godoy; Martins, Daniele dos Santos

doi:10.3390/ani11030598

Cited by 3 publications

(7 citation statements)

References 135 publications

(186 reference statements)

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“…We found that the gene expression levels of OCT4, NANOG, c-KIT, DAZL, STRA8, BOULE, and PIWIL2 increased remarkably in the bovine testis from the fetal to adult stage. It has been reported that the OCT4 and NANOG expression expands from gonocytes in the neonatal testis to spermatocytes and round spermatids after sexual maturation, which may explain the increased expression levels found in the adult stage ( 25 , 26 ). In concordance with these results, we immunodetected NANOG across the spermatogenic epithelium of the fetal testis with a high intensity in the Sertoli and pre-spermatogonial cells.…”

Section: Discussionmentioning

confidence: 99%

“…In concordance with these results, we immunodetected NANOG across the spermatogenic epithelium of the fetal testis with a high intensity in the Sertoli and pre-spermatogonial cells. Even though the roles of FRAGILIS and c-KIT have not been deeply studied in cattle, the expression of both genes occurs early in development, where they activate important mediators of postnatal meiotic and post-meiotic GC ( 26 ). In addition to the increasing expression levels of DAZL in the bovine testis during development, our analyses immunodetected DAZL mainly in the peripheral areas of bovine fetal seminiferous tubules associated to Sertoli and pre-spermatogonial cells.…”

Section: Discussionmentioning

confidence: 99%

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Overexpression of DAZL, STRA8, and BOULE Genes and Treatment With BMP4 or Retinoic Acid Modulate the Expression of MSC Overexpressing Germ Cell Genes

et al. 2021

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In vitro gamete derivation from stem cells has potential applications in animal reproduction as an alternative method for the dissemination of elite animal genetics, production of transgenic animals, and conservation of endangered species. Mesenchymal stem cells (MSCs) may be suitable candidates for in vitro gamete derivation considering their differentiative capacity and their potential for cell therapy. Due to its relevance in gametogenesis, it has been reported that retinoic acid (RA) and bone morphogenetic protein (BMP) 4 are able to upregulate the expression of specific markers associated to the early stages of germ cell (GCs) differentiation in bovine fetal MSCs (bfMSCs). In the present study, we used polycistronic vectors containing combinations of GC genes DAZL, STRA8, and BOULE followed by exposure to BMP4 or RA to induce GC differentiation of bovine fetal adipose tissue-derived MSC (AT-MSCs). Cells samples at Day 14 were analyzed according to the expression of pluripotent genes NANOG and OCT4 and GC genes DAZL, STRA8, BOULE, PIWI, c-KIT, and FRAGILIS using Q-PCR. Fetal and adult testis and AT-MSCs samples were also analyzed for the expression of DAZL, STRA8, and NANOG using immunofluorescence. Increased gene expression levels in the adult testis and cell-specific distribution of DAZL, STRA8, and NANOG in the fetal testis suggest that these markers are important components of the regulatory network that control the in vivo differentiation of bovine GCs. Overexpression of DAZL and STRA8 in bi-cistronic and DAZL, STRA8, and BOULE in tri-cistronic vectors resulted in the upregulation of OCT4, NANOG, and PIWIL2 in bovine fetal AT-MSCs. While BMP4 repressed NANOG expression, this treatment increased DAZL and c-KIT and activated FRAGILIS expression in bovine fetal AT-MSCs. Treatment with RA for 14 days increased the expression of DAZL and FRAGILIS and maintained the mRNA levels of STRA8 in bovine fetal AT-MSCs transfected with bi-cistronic and tri-cistronic vectors. Moreover, RA treatment repressed the expression of OCT4 and NANOG in these cells. Thus, overexpression of DAZL, STRA8, and BOULE induced the upregulation of the pluripotent markers and PIWIL2 in transfected bovine fetal AT-MSCs. The partial activation of GC gene expression by BMP4 and RA suggests that both factors possess common targets but induce different gene expression effects during GC differentiation in overexpressing bovine fetal AT-MSCs.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Overexpression of DAZL, STRA8, and BOULE Genes and Treatment With BMP4 or Retinoic Acid Modulate the Expression of MSC Overexpressing Germ Cell Genes

et al. 2021

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“…The first step is establishment of the chromosomal sex and begins the differentiation process. Sexual differentiation continues in sequential order to gonadal and phenotypical differentiation throughout gestation as the fetus responds to the hormonal or molecular signaling for normal development ( 1 , 2 ). In canines, the initiation of gonadal sexual differentiation begins around day 35–40 in gestation for distinguishing cell lines for male or female morphology ( 1 ).…”

Section: Introductionmentioning

confidence: 99%

“…Sexual differentiation continues in sequential order to gonadal and phenotypical differentiation throughout gestation as the fetus responds to the hormonal or molecular signaling for normal development ( 1 , 2 ). In canines, the initiation of gonadal sexual differentiation begins around day 35–40 in gestation for distinguishing cell lines for male or female morphology ( 1 ). The gonadal ridge develops around day 30–35 in gestation and this undifferentiated ridge will migrate and transform according to specific inputs for female vs. male morphology.…”

Section: Introductionmentioning

confidence: 99%

“…The gonadal ridge develops around day 30–35 in gestation and this undifferentiated ridge will migrate and transform according to specific inputs for female vs. male morphology. The gonadal portion of sexual differentiation directs primordial germ cell and embryologic tissue toward determining gonadal sex as testes or ovaries ( 1 , 3 ). The sexual determination process concludes with the development of the concomitant genital for the external and internal phenotypic sex.…”

Section: Introductionmentioning

confidence: 99%

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Case Report: Disorder of Sexual Development in a Chinese Crested Dog With XX/XY Leukocyte Chimerism and Mixed Cell Testicular Tumors

et al. 2022

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A 10-year-old intact female Chinese Crested dog was presented for evaluation and further diagnostics due to persistent symptoms of vulvar swelling, vaginal discharge, and an 8-year history of acyclicity. At presentation, generalized hyperpigmentation and truncal alopecia were identified, with no aberrations of the female phenotype. Vaginal cytology confirmed the influence of estrogen at multiple veterinary visits, and hormonal screening of progesterone and anti-Mullerian hormone indicated gonadal presence. Based on findings from abdominal laparotomy and gonadectomy, the tissue was submitted for histopathology. Histopathologic evaluation identified the gonads to be abnormal testes containing multiple Sertoli and interstitial (Leydig) cell tumors. The histopathologic diagnosis of testes and concurrent normal external female phenotype in the patient lead to a diagnosis of a disorder of sexual development (DSD). Karyotype evaluation by conventional and molecular analysis revealed a two cell line chimeric pattern of 78,XX (80%) and 78,XY (20%) among blood leukocytes, as well as a positive PCR test for the Y-linked SRY gene. Cytogenetic analysis of skin fibroblasts revealed the presence of 78,XX cells exclusively, and PCR tests for the Y-linked SRY gene were negative in the hair and skin samples. These results are consistent with an XX/XY blood chimerism. This is one of the few case reports of a canine with the diagnosis of leukocyte chimerism with normal female phenotypic external genitalia. This case illustrates a distinct presentation for hormonally active Sertoli cell tumorigenesis and demonstrates surgery as a curative treatment option for clinically affected patients.

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