Gene expression of SMIT1, SMIT2, and HMIT were evaluated using a CFX96 touch detection system (Bio-Rad, USA).
The levels of HMIT, myo-inositol phosphate synthase (MIPS), myoinositol oxygenase (MIOX), and neural growth factor (NGF) protein expression were measured in the cytosolic fraction and the membrane fraction was used for the evaluation of the expression of SMIT1, SMIT2, and HMIT.
The membranes were then incubated overnight with the primary antibody for NGF, MIPS, MIOX (1:100, Santa Cruz Biotechnology[R], USA), SMIT1, SMIT2, HMIT (1:1000, GenOne Biotechnologies, Brazil), or [beta]-actin (internal loading control) (1:5000, Sigma, USA).
Remarkably, we have found for the first time that SMIT2 was not expressed in rat SN, and that HMIT was abundantly expressed in both the SN and the DRG (Figure 2).
No significant alterations of HMIT gene expression was observed in this model of STZ-induced diabetes.
In this tissue, there was no significant change in HMIT expression in STZ-induced diabetes.
Therefore, we hypothesized that the transport of myoinositol via both SMIT1 or 2 and HMIT is compromised during STZ-induced diabetes.