Changes in hypothalamic expression of the Lin28/let-7 system and related MicroRNAs during postnatal maturation and after experimental manipulations of puberty

[Abstract] Lin28 and Lin28b are related RNA-binding proteins that inhibit the maturation of miRNAs of the let-7 family and participate in the control of cellular stemness and early embryonic development. Considerable interest has arisen recently concerning other physiological roles of the Lin28/let-7 axis, including its potential involvement in the control of puberty, as suggested by genome-wide association studies and functional genomics. We report herein the expression profiles of Lin28 and let-7 members in the rat hypothalamus during postnatal maturation and in selected models of altered puberty. The expression patterns of c-Myc (upstream positive regulator of Lin28), mir-145 (negative regulator of c-Myc), and mir-132 and mir-9 (putative miRNA repressors of Lin28, predicted by bioinformatic algorithms) were also explored. In male and female rats, Lin28, Lin28b, and c-Myc mRNAs displayed very high hypothalamic expression during the neonatal period, markedly decreased during the infantile-to-juvenile transition and reached minimal levels before/around puberty. A similar puberty-related decline was observed for Lin28b in monkey hypothalamus but not in the rat cortex, suggesting species conservation and tissue specificity. Conversely, let-7a, let-7b, mir-132, and mir-145, but not mir-9, showed opposite expression profiles. Perturbation of brain sex differentiation and puberty, by neonatal treatment with estrogen or androgen, altered the expression ratios of Lin28/let-7 at the time of puberty. Changes in the c-Myc/Lin28b/let-7 pathway were also detected in models of delayed puberty linked to early photoperiod manipulation and, to a lesser extent, postnatal underfeeding or chronic subnutrition. Altogether, our data are the first to document dramatic changes in the expression of the Lin28/let-7 axis in the rat hypothalamus during the postnatal maturation and after different manipulations that disturb puberty, thus suggesting the potential involvement of developmental changes in hypothalamic Lin28/let-7 expression in the mechanisms permitting/leading to puberty onset.Ministerio de Economia y Competitividad; BFU 2008-00984Ministerio de Economia y Competitividad; BFU 2011-25021Junta de Andalucía; P08-CVI-03788United States. National Institutes of Health HD025123-ARRAUnited States. National Science Foundation; IOS1121691Instituto de Salud Carlos III; PI10/00088Xunta de Galicia; IN845B-2010/187Xunta de Galicia; 10CSA916014P

Loss-of-function mutations in PNPLA6 encoding neuropathy target esterase underlie pubertal failure and neurological deficits in Gordon Holmes syndrome

PubMedID: 25033069Context: Gordon Holmes syndrome (GHS) is characterized by cerebellar ataxia/atrophy and normosmic hypogonadotropic hypogonadism (nHH). The underlying pathophysiology of this combined neurodegeneration and nHH remains unknown.Objective: We aimed to provide insight into the disease mechanism in GHS.Methods: We studied a cohort of 6 multiplex families with GHS through autozygosity mapping and whole-exome sequencing.Results: We identified 6 patients from 3 independent families carrying loss-of-function mutations in PNPLA6, which encodes neuropathy target esterase (NTE), a lysophospholipase that maintains intracellular phospholipid homeostasis by converting lysophosphatidylcholine to glycerophosphocholine. Wild-type PNPLA6, but not PNPLA6 bearing these mutations, rescued a well-established Drosophila neurodegenerative phenotype caused by the absence of sws, the fly ortholog of mammalian PNPLA6. Inhibition of NTE activity in the LßT2 gonadotropecell line diminished LH response to GnRH by reducing GnRH-stimulated LH exocytosis, without affecting GnRH receptor signaling or LHß synthesis.Conclusion: These results suggest that NTE-dependent alteration of phospholipid homeostasis in GHS causes both neurodegeneration and impaired LH release from pituitary gonadotropes, leading to nHH. Copyright © 2014 by the Endocrine Society.National Science Foundation: IOS112169

Histomorfometria e histoquímica dos ovários, tubas e útero de ratas hipotireóideas em metaestro-diestro Histomorphometry and histochemistry of the ovaries, oviduct and uterus in hypothyroid rats in the metaestrus-diestrus

A foliculogênese ovariana foi estudada em ratas adultas Wistar, hipotireóideas na fase de metaestro-diestro. O hipotireoidismo foi induzido pela administração oral e diária de propiltiouracil (1mg/animal). As ratas eutireóideas receberam placebo. Após 120 dias de tratamento, foi colhido o plasma para dosagem de tiroxina livre, progesterona e estradiol após o que foram sacrificadas para colheita dos ovários, tubas e útero, para avaliação histomorfométrica e histoquímica. O hipotireoidismo reduziu significativamente o peso dos ovários e o número de folículos secundários e terciários e de corpos lúteos sem, no entanto, alterar a porcentagem de folículos atrésicos e o número de folículos primários e pré-ovulatórios. As células da granulosa dos folículos secundários das ratas hipotireóideas apresentavam núcleo pequeno com significativa redução do número de regiões organizadoras de nucléolo (NORs). Essas mudanças não alteraram os valores periféricos de estradiol e de progesterona. Houve redução significativa da espessura do endométrio, do número de glândulas endometriais e da altura do epitélio do infundíbulo.<br>Ovarian folicullogenesis was studied in adult Wistar rats. The animals were hypothyroid in the metaestrus/diestrus phase of the estrous cycle. Hypothyroidism was induced by oral treatment with 1mg/animal of propylthiouracil. Euthyroid rats were kept under the same experimental conditions receiving placebo. After 120 days under treatment, all animals were killed and plasma was taken to assessing free thyroxine, progesterone, and estradiol. The ovaries, tubes and uterus were sampled for histomorphometric and histochemistry evaluation. Hypothyroidism significantly reduced the weight of the ovaries and the number of secondary and tertiary follicles, and corpora luteum. The number of atretic follicles as well as primary and pre-ovulatory follicles was not affected. Granulosa cells in the secondary follicles of treated rats had small nuclei with a reduced number of AgNORs. Significant reduction of the endometrium thickness, number of endometrial glands and the infundibulum epithelium height was observed