Comprehensive Analysis of Neonatal versus Adult Unilateral Decortication in a Mouse Model Using Behavioral, Neuroanatomical, and DNA Microarray Approaches

Abstract

Previously, studying the development, especially of corticospinal neurons, it was concluded that the main compensatory mechanism after unilateral brain injury in rat at the neonatal stage was due in part to non-lesioned ipsilateral corticospinal neurons that escaped selection by axonal elimination or neuronal apoptosis. However, previous results suggesting compensatory mechanism in neonate brain were not correlated with high functional recovery. Therefore, what is the difference among neonate and adult in the context of functional recovery and potential mechanism(s) therein? Here, we utilized a brain unilateral decortication mouse model and compared motor functional recovery mechanism post-neonatal brain hemisuction (NBH) with adult brain hemisuction (ABH). Three analyses were performed: (1) Quantitative behavioral analysis of forelimb movements using ladder walking test; (2) neuroanatomical retrograde tracing analysis of unlesioned side corticospinal neurons; and (3) differential global gene expressions profiling in unlesioned-side neocortex (rostral from bregma) in NBH and ABH on a 8 × 60 K mouse whole genome Agilent DNA chip. Behavioral data confirmed higher recovery ability in NBH over ABH is related to non-lesional frontal neocortex including rostral caudal forelimb area. A first inventory of differentially expressed genes genome-wide in the NBH and ABH mouse model is provided as a resource for the scientific community.

Two-color Dye-swap DNA Microarray approach toward confident gene expression profiling in PMCAO mouse model for ischemia-related and PACAP38-influenced genes

“…The samples were individually ground in liquid nitrogen to prepare very fine powders for subsequent downstream gene expression analyses [2] , [3] , [4] , [5] , [6] and proteomic analysis as required [4] . Prior to each DNA microarray analysis, the total RNA, extracted using an optimized protocol based on the QIAGEN RNeasy Mini Kit (QIAGEN, Germantown, MD, USA) quantity and quality was measured spectrophotometrically with NanoDrop (Thermo Scientific, Wilmington, DE, USA) and re-confirmed using formaldehyde-agarose gel electrophoresis [2] , [3] , [4] , [5] , [6] , [8] , [9] . Briefly, the obtained total RNA quality measurements revealed good quality as demonstrated by the A 260/280 values > 1.8, A 260/230 > 1.8; in our experiments, we always aim for a higher value of more than 2.0 to 2.3 [2] , [3] , [4] , [5] , [6] .…”

“…All the steps followed in sequence provided confidence that we had the best quality total RNA for use in the DNA microarray chip. As a last step, the Agilent mouse whole genome 4 × 44 K (G4122F) DNA slide (composed of 4 chips on one slide) was used for the microarray analysis, which was performed according to the Agilent instructions (Agilent Technologies, Santa Clara, CA, USA) and detailed in our publications [2] , [3] , [4] , [5] , [6] , [8] , [9] . As also mentioned below, our DNA microarray analysis experiment is based on the two-color dye-swap approach [10] .…”

Pituitary adenylate cyclase‐activating polypeptide promotes eccrine gland sweat secretion

“…Functional sweat‐secreting glands were visualized by the alternative Minor starch–iodine test as described previously . Adult C57BL/6J mice ( n = 10) were anaesthetized with 10 μL g −1 B.W.…”

“…Total RNA extraction and reverse transcriptase-polymerase chain reaction (RT-PCR) were performed as described previously. [24][25][26][27] Tissue samples from dome-shaped footpads of mice and from human plantar skin were obtained for RNA extraction. Following removal of the epidermis, dermis richtissues were used for mouse samples.…”

“…Functional sweat-secreting glands were visualized by the alternative Minor starch-iodine test as described previously. [24][25][26][27]30,31 Adult C57BL/6J mice (n = 10) were anaesthetized with 10 lL g À1 B.W. pentobarbital (Kyoritsu, Tokyo, Japan), and their paws coated with 10% povidoneiodine solution (Hakuzo Medical, Osaka, Japan).…”