Abstract

Laminin N-terminus α31 (LaNt α31) is a netrin-like protein derived from alternative splicing of the laminin α3 gene. Although LaNt α31 has been demonstrated to influence corneal and skin epithelial cell function, its expression has not been investigated beyond these tissues. In this study, we used immunohistochemistry to characterise the distribution of this protein in a wide-array of human tissue sections in comparison to laminin α3. The data revealed widespread LaNt α31 expression. In epithelial tissue, LaNt α31 was present in the basal layer of the epidermis, throughout the epithelium of the digestive tract, and in much of the epithelium of the reproductive system. LaNt α31 was also found throughout the vasculature of most tissues, with enrichment in reticular-like fibres in the extracellular matrix surrounding large vessels. A similar matrix pattern was observed around the terminal ducts in the breast and around the alveolar epithelium in the lung, where basement membrane staining was also evident. Specific enrichment of LaNt α31 was identified in sub-populations of cells of the kidney, liver, pancreas, and spleen, with variations in intensity between different cell types in the collecting ducts and glomeruli of the kidney. Intriguingly, LaNt α31 immunoreactivity was also evident in neurons of the central nervous system, in the cerebellum, cerebral cortex, and spinal cord. Together these findings suggest that LaNt α31 may be functionally relevant in a wider range of tissue contexts than previously anticipated, and the data provides a valuable basis for investigation into this interesting protein.

Highlights

  • Laminins (LMs) are essential extracellular matrix (ECM) structural proteins required for the assembly and function of basement membranes (BMs) [1, 2]

  • The C-terminus of Laminin N-terminal (LaNt) α31 is unique to this protein and not conserved with any human LMs or netrins (Fig 1A and 1B)

  • Previous publications have validated two separate antibodies that recognise human LaNt α31; rabbit polyclonal antibodies raised against the entire unique region of human LaNt α31 [6], and mouse monoclonal antibodies raised against a peptide within the LaNt α31-specific region [7] (Fig 1A and 1B)

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Summary

Introduction

Laminins (LMs) are essential extracellular matrix (ECM) structural proteins required for the assembly and function of basement membranes (BMs) [1, 2]. LMs have been studied extensively over the past 40+ years and 12 LM encoding genes have been identified in higher organisms [5]; the complexity of the family has further grown with the identification of a series of transcripts encoding non-laminin proteins that are generated from laminin-encoding genes by alternative splicing. LAMA3 is unique within the LM family in that is has two promoters giving rise to structurally distinct isoforms; a short LMα3a form and a longer LMα3b. These proteins share common carboxyl-terminal (C-terminal) regions but differ in the length of their amino terminus (N-terminus) [9, 10]. We used LaNt α31 specific monoclonal antibodies to determine the protein distribution across a wide range of human adult tissues for the first time, while comparing to total LMα3 localisation

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