ABSTRACT
In brief, we report an Iranian family with a history of both azoospermia and premature ovarian insufficiency with the same heterozygote mutation in the NR5A1 gene that can be transmitted. As far as we know, this is the first observation that a common mutation in NR5A1 can cause these above-mentioned phenotypes in a family.
Subject(s)
Azoospermia/genetics , Primary Ovarian Insufficiency/genetics , Steroidogenic Factor 1/genetics , Adult , Female , Heterozygote , Humans , Iran , Male , Mutation , PedigreeABSTRACT
We have cloned the PERF 15 gene, which encodes the most abundant protein of the perinuclear theca of rat spermatozoa. PERF 15 is related to the superfamily of lipophilic transport proteins. It has a molecular weight of 15,060 and is present exclusively in the subacrosomal region of the sperm head. Northern blot analysis demonstrated that this gene transcribed PERF 15 mRNA in meiotic and postmeiotic cells. The PERF 15 gene contains four exons and three introns. Exon 1 codes for amino acids 1-24, exon 2 for amino acids 25-82, exon 3 for amino acids 83-116, and exon 4 for amino acids 117-132. The three introns are composed of 2241, 547, and 164 base pairs (bp), respectively. The exon/intron boundaries are identical to those found in the mouse myelin P2 gene, but there is no resemblance in size and sequence between the corresponding introns of the PERF 15 and myelin P2 genes. Localization of the initiation transcription site by primer extension showed that the 5'-untranslated region of this gene is 67 bp upstream of the translation initiation site. Primer extension analysis also suggests that there is one transcription start site for this gene. Inverse polymerase chain reaction generated a 204-bp fragment, located upstream of the translation initiation codon, that has some homology with regions of other mammalian genes.
Subject(s)
Carrier Proteins/genetics , Spermatozoa/metabolism , Testicular Hormones/genetics , Animals , Base Sequence , Carrier Proteins/chemistry , Cloning, Molecular , DNA Primers/genetics , Exons , Fatty Acid-Binding Proteins , Genomic Library , Introns , Male , Mice , Molecular Sequence Data , Molecular Weight , Myelin P2 Protein/genetics , Polymerase Chain Reaction , RNA, Messenger/genetics , RatsABSTRACT
The perinuclear theca (PT) is a cytoskeletal structure that covers the nucleus of mammalian spermatozoa and is believed to have a membrane-binding role. The objectives of this study were to analyze the protein composition of the mouse PT, to identify and sequence its major protein component, and to characterize this protein's transcriptional and translational origins during spermatogenesis. The PT was extracted from demembranated and acrosome-depleted mouse sperm heads by alkaline treatment. The protein profile of the PT extract was composed of several polypeptides, of which a 15-kDa subacrosomal protein predominated and was found to be immunocross-reactive with a previously cloned 15-kDa PT protein of the rat (PERF 15) that belongs to a family of lipid-binding proteins. A primer pair designed from rat PERF 15 cDNA was then used to screen a mouse testicular cDNA library by polymerase chain reaction (PCR). The deduced amino acid sequence obtained from the PCR product was almost identical to the testicular-specific rat PERF 15. Developmental Northern blots and in situ hybridization studies performed with riboprobes encoding the mouse PERF 15 cDNA revealed that mRNA levels were highest in round and early elongating mouse spermatids. Immunohistochemistry indicated that PERF 15 began to be expressed in the cytoplasm of mid-pachytene spermatocytes and appeared to reach maximum expression in the distal cytoplasm of late elongating mouse spermatids, long after transcriptional arrest. During the development of round and early elongated spermatids, the immunolabel became progressively concentrated over the anterior half of the spermatid nucleus, suggesting a subacrosomal deposition of PERF 15 during this phase of mouse spermiogenesis.
