J Vet Sci.  2013 Dec;14(4):373-380. 10.4142/jvs.2013.14.4.373.

Time-dependent changes of calbindin D-28K and parvalbumin immunoreactivity in the hippocampus of rats with streptozotocin-induced type 1 diabetes

• Yi SS

• KMID: 1712302
• DOI: http://doi.org/10.4142/jvs.2013.14.4.373

Abstract

The hippocampus is affected by various stimuli that include hyperglycemia, depression, and ischemia. Calcium-binding proteins (CaBPs) have protective roles in the response to such stimuli. However, little is known about the expression of CaBPs under diabetic conditions. This study was conducted to examine alterations in the physiological parameters with type 1 diabetes induced with streptozotocin (STZ) as well as time-dependent changes in the expression of two CaBPs changes of were being evaluated. Rats treated with STZ (70 mg/kg) had high blood glucose levels (>21.4 mmol/L) along with increased food intake and water consumption volumes compared to the sham controls. In contrast, body weight of the animals treated with STZ was significantly reduced compared to the sham group. CB-specific immunoreactivity was generally increased in the hippocampal CA1 region and granule cell layer of the dentate gyrus (DG) 2 weeks after STZ treatment, but decreased thereafter in these regions. In contrast, the number of PV-immunoreactive neurons and fibers was unchanged in the hippocampus and DG 2 weeks after STZ treatment. However, this number subsequently decreased over time. These results suggest that CB and PV expression is lowest 3 weeks after STZ administration, and these deficits lead to disturbances in calcium homeostasis.

Keyword

calbindin D-28K; calcium binding protein; hippocampus; parvalbumin; type I diabetes

MeSH Terms

Animals
Calbindin 1/*genetics/metabolism
Diabetes Mellitus, Experimental/*chemically induced
Diabetes Mellitus, Type 1/*chemically induced
*Gene Expression Regulation
Hippocampus/*metabolism
Male
Parvalbumins/*genetics/metabolism
Rats
Rats, Wistar
Streptozocin/administration & dosage
Calbindin 1
Parvalbumins
Streptozocin

Figure

• 

  Fig. 1 Physiological data for the sham and streptozotocin (STZ)-treated rats. Body weights of the sham group exponentially increased while those of the STZ-treated rats slowly decreased over the 4-week study period (A). Blood glucose levels at 0, 1, 2, 3, and 4 weeks after STZ treatment (Pre, STZ1w, STZ2, STZ3w, and STZ4w, respectively) were significantly different from those of the sham controls. Blood glucose levels continuously increased in the STZ-treated animals from 21.4 mmol/L 1 week after STZ administration (B). Food intake (C) and water consumption (D) also increased 1 week after STZ treatment. *p < 0.005.

• 

  Fig. 2 Immunohistochemical staining for calbindin D-28K (CB) in the hippocampus of sham and STZ-treated rats. The whole hippocampus can be seen in panels A, E, I and M, and the images of the CA1, CA2/CA3, and dentate gyrus (DG) are outlined by black squares. Neurons in the stratum pyramidale were rarely positive in the CA1 regions of the sham controls (B) whereas immunoreactivity observed 2~4 weeks after STZ treatment was stronger (F, J and N). Few CB-positive neurons were observed in the stratum pyramidale of the CA2/CA3; however, CB-specific immunoreactivity in the mossy fibers was strong in all groups (C, G, K and O). Quantification of immunoreactivity in the mossy fibers and granule cell fibers is presented in Table 2. The number of CB-positive cells was decreased 3 weeks after STZ treatment and increased at 4 weeks. Some CB-positive neurons in the CA1 and CA2/CA3 are indicated by black arrowheads in panels F, G, J, K and O. A few CB-positive neurons in the granular cell layer (GCL) of the DG are indicated by black arrowheads in panel H. Time-dependent changes in CaBP-positive staining according to hippocampal region are described in Table 1. GCL: granular cell layer, MF: mossy fiber, ML: molecular layer, PL: polymorphic layer, SO: stratum oriens, SR: stratum radiatum, SP: stratum pyramidale. Scale bars = 100 µm.

• 

  Fig. 3 Immunohistochemical staining for parvalbumin (PV) in the hippocampus of sham and STZ-treated rats. The whole hippocampus can be seen in panels A, E, I and M while images of the CA1, CA2/CA3, and DG are outlined by black squares. PV-positive neurons in the CA1 were observed in the stratum oriens, stratum radiatum, and stratum pyramidale, but were primarily found in the stratum pyramidale (panels B, F, J and N). PV-positive neurons were also found along with the granular cell layer and polymorphic layer of the DG. A few PV-positive neurons in the CA1 and CA2/CA3 are indicated by black arrowheads in panels B, C, F, G, J, K and O. Some CB-positive neurons in the DG are indicated by black arrowheads in panels D, H, L and P. Immunoreactivity specific for PV-positive neurons in the CA1 and CA2/CA3 regions gradually decreased over time (Table 1). Staining in the DG was first significantly reduced 3 weeks after STZ treatment and then rose to the same level found in the sham control at 4 weeks. Scale bars = 100 µm.

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