**3. Comparison of sequences and construction of phylogenic tree of avian muscarinic subsets**

Multiple sequence alignment was done with NCBI tool (https://www.ncbi.nlm. nih.gov/blast/) and individual phylogenic tree for each receptor among avian were created using MEGA 11. Respective human receptors were used for comparative purpose (**Figures 2**–**4**) similarity between the reported avian sequences as shown in

#### **Figure 2.**

*Phylogeny tree prepared based on sequence similarity of mAChR 2 of C. livia with different species prepared using MEGA11.*

*Avian Muscarinic Receptors: An Update DOI: http://dx.doi.org/10.5772/intechopen.111720*

the tables (**Tables 1**–**3**). By analyzing **Table 1** the percentage identity of mAChR 2 of *Columba livia* (rock pigeon) with that of *Aquila chrysaetos* (golden eagle) is 95.5% with maximum percentage similarity and minimum similarity with cholinergic muscarinic receptor 2 of *Homo sapiens* (human beings). Similarly, **Table 3** details gives percentage identity of muscarinic receptor 3 with other species and its respective human receptors. The same applies to **Table 4** and **Figure 4**.

#### **Figure 3.**

*Phylogeny tree prepared based on sequence similarity of mAChR 3 of C. livia with different species prepared using MEGA.*

#### **Figure 4.**

*Phylogeny tree prepared based on sequence similarity of mAChR 4 of C.livia with different species prepared using MEGA11.*

*Avian Muscarinic Receptors: An Update DOI: http://dx.doi.org/10.5772/intechopen.111720*


#### **Table 1.**

*Sequence similarity of mAChR 2 of C. livia with different species. Data collected from pubmed (https://www. ncbi.nlm.nih.gov/) and created with NCBI-nucleotide-BLAST (https://www.ncbi.nlm.nih.gov/blast/).*



#### **Table 2.**

*Sequence similarity of mAChR 3 of C. livia with different species. Data collected from pubmed (https://www. ncbi.nlm.nih.gov/) and created with NCBI-nucleotide-BLAST (https://www.ncbi.nlm.nih.gov/blast/).*



#### **Table 3.**

*Sequence similarity of mAChR 4 of C. livia with different species. Data collected from pubmed (https://www. ncbi.nlm.nih.gov/) and created with NCBI-nucleotide-BLAST (https://www.ncbi.nlm.nih.gov/blast/).*



#### **Table 4.**

*Distribution of subtypes of muscarinic receptors in various species and their location.*
