Localization of Fertility Factor SP22 to Specific Cell Types Within the Anterior Pituitary Gland

Abstract

Sperm protein 22 (SP22) was recently identified in the anterior pituitary gland (AP) of male Golden Syrian hamsters using ion trap mass spectrometry. SP22 has been implicated in apoptosis, androgen receptor function, fertility, and ontogeny of early-onset Parkinson's disease. However, the role of SP22 in the pituitary has not been investigated. We cloned the cDNA for full-length SP22 from AP and posterior lobe (posterior pituitary and intermediate lobe) of the pituitary gland in adult male rats and Golden Syrian hamsters, confirming the presence of SP22 mRNA in the AP and posterior lobe. Because gonadal steroids are important regulators of AP function, and SP22 is associated with androgen receptor function, we used Western blots to compare SP22 in the AP of intact and orchidectomized male rats given placebo or a low or high dose of testosterone. SP22 did not differ with treatment, indicating that AP SP22 concentration was not regulated by testosterone. To localize SP22 to specific cells of the AP, mirror-image paraffin sections were labeled against SP22 and either luteinizing hormone (LH)beta, thyroid-stimulating hormone (TSH)beta, prolactin, adrenocorticotropic hormone (ACTH), or growth hormone (GH) using peroxidase-conjugated secondary antibody. Additional sections were colabeled with SP22 and one of the AP hormones using fluorescent secondary antibodies. SP22 colocalized in somatotropes and thyrotropes in rat and hamster. We identified SP22 in a small percentage of corticotropes, gonadotropes, and lactotropes. This is the first report that SP22 mRNA is present specifically in the AP, and SP22 is localized primarily in somatotropes and thyrotropes. SP22 may help regulate AP function and be particularly important for the control of GH and TSH secretion.

Proteomics of the Anterior Pituitary Gland as a Model for Studying the Physiology of a Heterogeneous Organ

“…Amplicons were subcloned into plasmids and sequenced to confirm the identity of the SP22 cDNA. DNA sequencing confirmed the presence of the SP22 mRNA amplified in the AP and posterior lobe of the hamster and rat (34).…”

“…We identified SP22 in a small percentage of corticotropes, gonadotropes, and lactotropes. This was the first report that SP22 was present specifically in the AP and localized primarily in somatotropes and thyrotropes, suggesting that SP22 may help regulate AP function and be particularly important for the control of GH and TSH secretion (34).…”

“…The RNA was extracted and used for reverse transcriptase-polymerase chain reaction (RT-PCR). Total RNA (l ug) from AP, posterior lobe, and testis (positive control) was reverse transcribed using MuLV reverse transcriptase and amplified using Taq DNA polymerase (34). Primers were made using known homologous sequences for hamster and rat SP22 available in GenBank.…”